Low-frequency Stimulated Research Articles

Low-Frequency Stimulated Light Scattering in Virus Suspensions under Picosecond Excitation

Low-Frequency Stimulated Light Scattering in Virus Suspensions under Picosecond Excitation

Plasmonic Resonance of Low-Frequency Stimulated Raman Scattering during Water Optical Breakdown

Plasmonic Resonance of Low-Frequency Stimulated Raman Scattering during Water Optical Breakdown

Low-Frequency Stimulated Scattering in an Aqueous Suspension of Dielectric Nanospheres

A low-frequency stimulated scattering (LFSS) signal caused by pulsations of nanospheres under an electromagnetic field is detected in an aqueous suspension of dielectric nanospheres of three diameters (∼74 nm, ∼292 nm, and ∼540 nm) at various particle concentrations. The frequency shifts of the scattering line, calculated based on the developed model, are in good agreement with the measurement results.

Low-Frequency Stimulated Raman Scattering on Acoustic Vibrations of Dielectric Nanospheres in Aqueous Suspension

A low-frequency SRS (LFSRS) signal is detected at a certain dielectric nanoparticle (latex spheres) concentration in an aqueous suspension. The SRS line shift is close to its estimate by the “transverse” hypersound speed in latex.

The effects of therapeutic electric stimulation on acute muscle atrophy in rats after spinal cord injury

Misawa A, Shimada Y, Matsunaga T, Sato K. The effects of therapeutic electric stimulation on acute muscle atrophy in rats after spinal cord injury. Arch Phys Med Rehabil 2001;82:1596-603. Objective: To evaluate the effects of electric stimulation in preventing acute muscle atrophy after spinal cord transection in rats. Design: A randomized experimental design. Setting: Animal facilities for experimental medicine. Animals: Fifty-six adult male Wistar rats assigned to control, low-frequency, and high-frequency groups. Interventions: The rats were implanted with a percutaneous intramuscular electrode in the vicinity of the peroneal nerve; then the spinal cord was transected in a T9 level. The stimulation frequency was low (20Hz) or high (100Hz). The stimulation cycle was 4 seconds of stimulation every 8 seconds. Main Outcome Measurements: The lesser fiber diameters from type 1, 2A, and 2B muscle fibers were measured. In another assessment, maximal contraction force was measured. The muscle force produced at 20 and 100Hz was expressed as increasing values in tetanic force. Results: Comparison between nonstimulated and stimulated tibialis anterior muscles found that atrophy of type 1 fibers (p < .01) and type 2B fibers (p < .05) at both stimulated levels and of type 2A fibers at 100-Hz level (p < .05) was prevented by therapeutic electric stimulation (TES). There were significant differences in the size of muscle fiber diameter between nonstimulated and stimulated muscles at 100Hz in type 2A and, markedly, in type 2B. The increasing value of muscle force was significantly greater at 100Hz than at 20Hz (p < .05). No significant histologic differences were observed between high- and low-frequency stimulated fibers of any of the 3 muscle types. Conclusions: Acute atrophy of muscle fibers was more effectively prevented by high-frequency stimulation (100Hz) than by no stimulation or low-frequency stimulation (20Hz). The increasing value of muscle force was significantly greater at high-frequency than low-frequency stimulation, suggesting that the clinical application of high-frequency stimulation in acute spinal cord injury should be studied. © 2001 by the American Congress of Rehabilitation Medicine and the American Academy of Physical Medicine and Rehabilitation

Inactivation of sarcoplasmic reticulum Ca(2+)-atpase in low-frequency stimulated rat muscle.

Continuous low-frequency stimulation (CLFS) by implanted electrodes for 12-24 h led to a significant (approximately 30%) decrease in the activity of sarcoplasmic reticulum Ca(2+)-ATPase in fast-twitch extensor digitorum longus (EDL) and tibialis anterior (TA) muscles of intact rats. The decline in catalytic activity after 24 h of CLFS was accompanied by an approximately twofold increase in dinitrophenylhydrazine-reactive carbonyl groups of the enzyme. It also correlated with an immunochemically determined 30% decrease in Ca2(+)-ATPase protein. Recovery studies after 12 h of CLFS revealed a relatively slow (48-72 h) re-establishment of normal catalytic activity. These findings suggest that the 30% decline of Ca(2+)-ATPase activity in low-frequency stimulated rat muscle led to an irreversible modification by protein oxidation. The decrease in Ca(2+)-ATPase protein most likely resulted from the degradation of inactive Ca(2+)-ATPase molecules. The relatively slow recovery of Ca(2+)-ATPase activity suggests that de novo synthesis of the enzyme may be necessary to re-attain normal activity.

Calcium transients in single fibers of low-frequency stimulated fast-twitch muscle of rat.

Ca(2+) transients were investigated in single fibers isolated from rat extensor digitorum longus muscles exposed to chronic low-frequency stimulation for different time periods up to 10 days. Approximately 2.5-fold increases in resting Ca(2+) concentration ([Ca(2+)]) were observed 2 h after stimulation onset and persisted throughout the stimulation period. The elevated [Ca(2+)] levels were in the range characteristic of slow-twitch fibers from soleus muscle. In addition, we noticed a transitory elevation of the integral [Ca(2+)] per pulse with a maximum ( approximately 5-fold) after 1 day. Steep decreases in rate constant of [Ca(2+)] decay could be explained by an immediate impairment of Ca(2+) uptake and, with longer stimulation periods, by an additional loss of cytosolic Ca(2+) binding capacity resulting from a decay in parvalbumin content. A partial recovery of the rate constant of [Ca(2+)] decay in 10-day stimulated muscle could be explained by an increasing mitochondrial contribution to Ca(2+) sequestration.

Protein oxidation, tyrosine nitration, and inactivation of sarcoplasmic reticulum Ca 2+-ATPase in low-frequency stimulated rabbit muscle

Sustained contractile activity by chronic low-frequency stimulation in rabbit fast-twitch muscle causes a partial (40–50%) inactivation of the sarcoplasmic reticulum (SR) Ca 2+-ATPase and, with prolonged stimulation, a SERCA1a to SERCA2a transition. To investigate the underlying mechanism of the inactivation which precedes the isoform transition, we analyzed SR from 4-day stimulated muscles for Ca 2+-ATPase activity, lipid peroxidation, SH and carbonyl groups, and nitrotyrosine. At unaltered SH group and malondialdehyde contents, carbonyl groups were elevated 50% in the SR from stimulated muscles. Immunoblotting with anti-dinitrophenyl and anti-nitrotyrosine antibodies revealed strong labeling of the Ca 2+-ATPase, suggesting the inactivation of the enzyme to result from protein oxidation and peroxynitrite-mediated tyrosine nitration.

Fiber transformation and replacement in low-frequency stimulated rabbit fast-twitch muscles.

The fast-to-slow conversion of rabbit skeletal muscles by chronic low-frequency (10 Hz, 12 h daily) stimulation involves (1) sequential fast-to-slow fiber-type transitions in the order of type IID-->type IIA-->type I, and (2) the replacement of deteriorating fast-twitch glycolytic fibers by new fibers derived from satellite cells and myotubes. These two processes were analyzed in 30- and 60-day stimulated extensor digitorum longus and tibialis anterior muscles. Fast-to-slow transforming fibers were identified by myofibrillar actomyosin histochemistry as type C fibers and immunohistochemically by their reaction with monoclonal antibodies specific to slow and fast myosin heavy chain isoforms. In situ hybridization of mRNA specific to the myosin heavy chain I isoform identified all fibers expressing slow myosin, i.e., type I and C fibers. The fraction of transforming fibers ranged between 35% and 50% in 30-day stimulated muscles. The percentage of type I fibers (20%) was threefold elevated in extensor digitorum longus muscle, but unaltered (3.5%) in tibialis anterior muscle, suggesting that fast-to-slow fiber conversion was more advanced in the former than in the latter. Fiber replacement was indicated by the finding that the fiber populations of both muscles contained 15% myotubes or small fibers with central nuclei. In situ hybridization revealed that myotubes and small regenerating fibers uniformly expressed myosin heavy chain I mRNA. Similarly, high percentages of slow-myosin-expressing myotubes and small fibers were found in 60-day stimulated muscles.

Reverse transcriptase-polymerase chain reaction detects induction of cardiac-like α myosin heavy chain mRNA in low frequency stimulated rabbit fast-twitch muscle

Using reverse transcriptase-polymerase chain reaction we quantified in rabbit skeletal muscles expression levels of the highly homologous cardiac α and β myosin heavy chain (αMHC, βMHC) mRNA isoforms. Masseter muscle displayed highest levels of a cardiac-like αMHC mRNA. This isoform was present at 20-fold lower amounts in slow soleus and at 200-fold lower levels in several fast-twitch muscles. Low-frequency stimulation periods exceeding 20 days drastically induced the αMHC mRNA in fast tibialis anterior. The αMHC mRNA was 140-fold elevated after 60 days when βMHC mRNA had increased 50-fold. Our results demonstrate the wide distribution of a cardiac-like αMHC mRNA in skeletal muscle and its marked induction during fast-to-slow transition as induced by low-frequency stimulation.

Low-frequency stimulation of rat fast-twitch muscle enhances the expression of hexokinase II and both the translocation and expression of glucose transporter 4 (GLUT-4).

This study followed changes in the capacities of uptake and phosphorylation of glucose in response to contractile activity in low-frequency stimulated (10Hz, 24 h/d) rat fast-twitch muscle. We investigated the intracellular distribution of GLUT-4, the major glucose transporter isoform in muscle, changes in the amounts of its specific mRNA and total cellular protein, as well as changes in its relative synthesis rate. These analyses were complemented by measurements of total hexokinase activity and hexokinase II (HKII) expression at the levels of mRNA content and protein synthesis. Changes in protein synthesis were determined by in vivo labeling with [35S]methionine. Translocation of GLUT-4 into the sarcolemma was an immediate response to contractile activity, whereas changes in its total amount were observed only with ongoing stimulation (5 d and longer). A twofold increase in GLUT-4 content after 5 d and longer stimulation periods was preceded by elevations of its mRNA and by enhanced [35S]methionine incorporation. Conversely, increases in HKII expression with a rise in total hexokinase activity occurred soon after the onset of stimulation (30-fold elevations of HKII mRNA after 12 h and 20-fold increases in [35S]methionine incorporation after 24 h). With ongoing stimulation, HKII mRNA and synthesis returned to lower levels (fivefold elevations). Nevertheless, hexokinase activity continued to rise, stabilizing at fivefold-elevated levels after 3 d. These observation suggested that posttranscriptional mechanisms contributed to the upregulation of HKII, e.g. stabilization by elevated intracellular glucose and mitochondrial binding of the enzyme. This suggestion was supported by experiments with cessation after 24 h where hexokinase activity continued to increase, although the mRNA content and, especially, the [35S]methionine incorporation decayed steeply. The increase in HKII prior to GLUT-4 suggests that phosphorylation may be rate limiting in glucose utilization of glycolytic fibers under conditions of sustained contractile activity. Taken together, the changes in distribution and content of GLUT-4, as well as in HKII represent early metabolic adaptations. In addition, they are related to the overall process of stimulation-induced fiber type transformation.

Altered glucose 1,6-bisphosphate and fructose 2,6-biphosphate levels in low-frequency stimulated rabbit fast-twitch muscle

Glucose 1,6-bisphosphate (Glc-1,6-P 2) and fructose 2,6-bisphospate (Fru-2,6-P 2) concentration display pronounced increases in rabbit fast-twitch muscle during chronic low-frequency stimulation. These increases are first seen after stimulation periods exceeding 3 h and reach maxima after 12–24h of stimulation (approximately 3-fold for Glc-1,6-P 2 and 5-fold for Fru-2,6-P 2). Both matabolites regress to normal values after stimulation periods longer than 4 days. The fact that their increases coincide with the replenishment of glycogen after its initial depletion, could point to a role of Glc-1,6-P 2 and Fru-2,6-P2 in glycogen metabolism.

Effect of electrical stimulation on the viability of the hippocampal slice preparation

Continuous electrical stimulation of rat hippocampal slices at a frequency of 1 Hz brought about a 50% decline in the evoked population spike amplitude at a rate 5 times faster than that caused by very low frequency ( 1 600 Hz) stimulation. Within 2 hr after the high frequency stimulation began the evoked response totally disappeared. By contrast low frequency stimulated slices maintained an evoked response for at least 9 hr. Continuous electrical stimulation, especially at high frequency seems to facilitate the deterioration of the in vitro hippocampal slice preparation.

Met5-enkephalin-Arg6-Phe7 inhibition of noradrenaline and acetylcholine release from peripheral organs.

The effect of the opioid heptapeptide Met5-enkephalin-Arg6-Phe7 was measured on the contractions of the longitudinal muscle of the guinea-pig ileum and mouse vas deferens; on release of endogenous and newly synthesized acetylcholine from the longitudinal muscle strip preparation of the guinea-pig ileum and of 3H-noradrenaline from mouse vas deferens. Met5-enkephelin-Arg6-Phe7 depressed the contractions of the longitudinal muscle strip of guinea-pig ileum elicited by low frequency stimulation (IC50 366.5 +/- 71.5 nM). The Ke value of naltrexone against the heptapeptide was found to be 0.50 +/- 0.08 nM. Bestatin prolonged and strongly potentiated the action of the opioid peptide in this tissue while captopril proved to be only slightly effective. Met5-enkephalin-Arg6-Phe7 also inhibited neuroeffector transmission in the mouse vas deferens with an IC50 value of 9.44 +/- 2.24 nM. The Ke value of naltrexone was 6.33 +/- 1.22 nM against the heptapeptide. Met5-enkephalin-Arg6-Phe7 (10(-5) M) failed to affect the low frequency stimulated release of endogenous acetylcholine from the guinea-pig ileum preparation. Addition of either bestatin or captopril with the heptapeptide resulted in a reduction of acetylcholine output. The opioid peptide by itself decreased the release of newly synthesized acetylcholine outflow from this organ. Met5-enkephalin-Arg6-Phe7 also reduced the 3H-noradrenaline outflow from mouse vas deferens and this effect was antagonized by naltrexone. From our data we conclude that Met5-enkephalin-Arg6-Phe7 affects the neural transmission of peripheral autonomically innervated organs by depressing the release of neurotransmitter. In vitro it has enkephalin-like character with a preference for the so called delta receptor interaction.